The study compares the isotopic niche (δ13C–δ15N) of three species of penaeid shrimps (Xiphopenaeus kroyeri, Artemesia longinaris and Litopenaeus schmitti) that are targets of fishing in south-eastern Brazil. The two hypotheses raised are based on the niche theory, in which the coexistence between organisms that have similar dietary demands is possible due to trophic partitioning: (i) shrimp species that share the habitat have segregated isotopic niches; and (ii) stages of maturity and genders vary with the trophic habitat (δ13C) and/or trophic position (δ15N). In multispecific fishing grounds, the isotopic niches of the shrimp species were segregated or had low overlap, indicating trophic partitioning. The intraspecific comparison of δ13C and δ15N showed a similar trend in all species. For δ13C, the values were more enriched in adult individuals than in juveniles, but similar between males and females. For δ15N, adult individuals also had more enriched values, and males were more enriched than females. The results confirm the hypotheses raised, and the assumptions of niche theory apply to these penaeid shrimps.